Gonzalez R, Arancibia R, Cáceres M, Martínez J, Smith P C
Institute of Nutrition and Food Technology, University of Chile, Santiago, Chile.
J Periodontal Res. 2009 Jun;44(3):386-94. doi: 10.1111/j.1600-0765.2008.01114.x. Epub 2009 Feb 6.
Tobacco smoking is a significant risk factor for periodontal disease. It has been suggested that smoking may alter connective tissue remodeling in the periodontium. In the present study, we investigated whether cigarette smoke condensate modulates the production of the serine protease urokinase in human gingival fibroblasts.
Primary cultures of human gingival fibroblasts were stimulated with cigarette smoke condensate. Urokinase production was evaluated through casein zymography and western blotting. Plasmin activation was assessed by means of a radial diffusion assay. The roles of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and reactive oxygen species in cigarette smoke condensate-stimulated urokinase production were studied using distinct selective inhibitors (SP600125, PD98059, N-acetyl cysteine). Reactive oxygen species production was determined using a fluorometric assay. Activation of ERK and JNK pathways were evaluated using western blots.
In gingival fibroblasts, cigarette smoke condensate potently stimulated urokinase production and plasmin activation. Cigarette smoke condensate-stimulated urokinase production was dependent on the activity of ERK/JNK pathways and was inhibited by the reactive oxygen species scavenger, N-acetyl cysteine. Cigarette smoke condensate strongly stimulated ERK and JNK phosphorylation and the generation of reactive oxygen species.
Cigarette smoke condensate stimulates urokinase production and plasmin activation in gingival fibroblasts. Moreover, cigarette smoke condensate-stimulated urokinase production depends on both the activation of ERK/JNK pathways and on the generation of intracellular reactive oxygen species. These results show that cigarette smoke may alter connective tissue remodeling by inducing production of the urokinase-type plasminogen activator through specific signaling pathways.
吸烟是牙周病的一个重要危险因素。有人提出，吸烟可能会改变牙周组织中的结缔组织重塑。在本研究中，我们调查了香烟烟雾冷凝物是否会调节人牙龈成纤维细胞中丝氨酸蛋白酶尿激酶的产生。
用人牙龈成纤维细胞的原代培养物用香烟烟雾冷凝物刺激。通过酪蛋白酶谱法和蛋白质印迹法评估尿激酶的产生。通过径向扩散试验评估纤溶酶激活。使用不同的选择性抑制剂（SP600125、PD98059、N-乙酰半胱氨酸）研究细胞外信号调节激酶（ERK）、c-Jun N端激酶（JNK）和活性氧在香烟烟雾冷凝物刺激的尿激酶产生中的作用。使用荧光测定法测定活性氧的产生。使用蛋白质印迹法评估ERK和JNK途径的激活。
在牙龈成纤维细胞中，香烟烟雾冷凝物强烈刺激尿激酶的产生和纤溶酶激活。香烟烟雾冷凝物刺激的尿激酶产生依赖于ERK/JNK途径的活性，并被活性氧清除剂N-乙酰半胱氨酸抑制。香烟烟雾冷凝物强烈刺激ERK和JNK磷酸化以及活性氧的产生。
香烟烟雾冷凝物刺激牙龈成纤维细胞中尿激酶的产生和纤溶酶激活。此外，香烟烟雾冷凝物刺激的尿激酶产生既依赖于ERK/JNK途径的激活，也依赖于细胞内活性氧的产生。这些结果表明，香烟烟雾可能通过特定信号通路诱导尿激酶型纤溶酶原激活剂的产生，从而改变结缔组织重塑。
